Abstract: Objective To detect pre-S1 antigen (HBV pre-S1-Ag), pre-S2 antigen (HBV pre-S2-Ag), hepatitis B virus (HBV) DNA and hepatitis B e ntigen (HBeAg)  in hepatitis B patients and discuss  the correlation among them and the clinical applications value. Methods Enzyme-liked immunosorbent adsorption method (ELISA) were applied  to detect HBVpre-S1-Ag, HBV pre-S2-Ag and markers of hepatitis B virus (HBV-M). Fluorescence quantitative polymerase couplet reaction method (FQ-PCR) was applied to detect HBV DNA and the detection results were analyzed statistically. Results Among the HBsAg positive patients, the pre-S1 antigen, pre-S2 antigen, HBV DNA positive patients were 594 cases, 541 cases, 629 cases, respectively, with the rate of 66.29%, 60.38%, 70.20%. HBeAg positive patients' pre-S1 antigens, pre-S2 antigens, HBV DNA positie rates were 90.21%, 74.46%, 93.32%, significantly higher than that in HBeAg negative patients with 45.28%,48.01% and 49.89%, respectively. The difference were statistically significant (P ＜ 0.01). Pre-S1-Ag, pre-S2-Ag and HBeAg positive rates arised with HBV DNA arising. Conclusions Pre-S1-Ag, pre-S2-Ag and HBV DNA and may contribute to hepatitis B early diagnosis and clinical observation and prognosis.